PXD046160

PXD046160 is an original dataset announced via ProteomeXchange.

Title	Residues 1 to 563 of α-hemolysin alone form a fully functional pore and their proper membrane insertion depends on the acylation status of the molecule
Description	The acylated pore-forming Repeats in ToXin (RTX) cytolysins α-hemolysin (HlyA) and adenylate cyclase toxin (CyaA) bind primarily to β2 integrins of leukocytes. HlyA binds the common CD18 subunit of the β2 integrins and CyaA selectively binds the CD11b subunit of complement receptor 3 (CR3). However, both toxins can also bind and permeabilize membranes of a variety of nonmyeloid cells. We constructed HlyA1-563/CyaA860-1706 hybrid molecules activated by the CyaA-activating acyltransferase CyaC, or by the HlyA-activating acyltransferase HlyC. We show that the C-terminal portion of the HlyA molecule, comprising the acylated segment and the RTX domain (residues 564 to 1024), can be functionally swapped with the CyaC-activated acylated segment bearing palmitoylated Lys983 and the much larger RTX domain of CyaA. Compared to the CD18-interacting HlyA, the CR3-interacting HlyA1-563/CyaA860-1706 hybrid exhibited a selectively reduced cytotoxicity on human THP-1 monocytes and Chinese hamster ovary (CHO) cells expressing CR3 (CHO-CR3) or lymphocyte function-associated antigen 1 (CHO-LFA1). However, the mono-palmitoylated HlyA1-563/CyaA860-1706 hybrid remained fully hemolytic and cytolytic toward erythrocytes and mock-transfected CHO cells and exhibited a comparable membrane activity on artificial planar lipid membranes as the bi-myristoylated HlyA. Thus, regardless of the length or number of the attached fatty acyl chains, the CyaC-activated HlyA1-563/CyaA860-1706 hybrid and the intact HlyA were comparably active on cells or artificial membranes lacking β2 integrins. These data suggest that once the hydrophobic pore-forming domain comprising N-terminal half of HlyA is brought into close contact with the cell membrane by the mono-palmitoylated acylated segment of CyaA, it can efficiently form fully cytolytic HlyA-like membrane pores.
HostingRepository	PRIDE
AnnounceDate	2024-03-25
AnnouncementXML	Submission_2024-03-25_08:12:33.855.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	David Jurnečka
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	palmitoylated residue; myristoylated residue
Instrument	Bruker Daltonics solarix series

Revision	Datetime	Status	ChangeLog Entry
0	2023-10-16 08:05:02	ID requested
⏵ 1	2024-03-25 08:12:34	announced

Dataset with its publication pending

submitter keyword: acylation,RTX toxin

Laboratory of Molecular Biology of Bacterial Pathogens
contact affiliation	Institute of Microbiology of the Czech Academy of Sciences, Prague, Czech Republic
contact email	sebo@biomed.cas.cz
lab head
David Jurnečka
contact affiliation	Institute of Microbiology, Czech Academy of Sciences
contact email	david.jurnecka@biomed.cas.cz
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/03/PXD046160

PRIDE project URI

• PRIDE
  1. PXD046160
     1. Label: PRIDE project
     2. Name: Residues 1 to 563 of α-hemolysin alone form a fully functional pore and their proper membrane insertion depends on the acylation status of the molecule